Retroviral RLI immunomodulatory gene therapy for glioblastoma

逆转录病毒 RLI 免疫调节基因治疗胶质母细胞瘤

基本信息

批准号：
10522026
负责人：
Manish Aghi
金额：
$ 40.38万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10522026
关键词：
Accounting Address Antigen Presentation Apoptosis Biological Availability Bone Marrow Brain Neoplasms CD8B1 gene CRISPR interference CRISPR screen CSF3 gene CTLA4 gene Cell physiology Cells Chimeric Proteins Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complex Custom Cytosine deaminase Cytotoxic Chemotherapy Data Diagnosis Enzymes Failure Gene Delivery Gene Expression Genes Glioblastoma Glioma IL7 gene Immune Immune Evasion Immune checkpoint inhibitor Immune response Immunocompetent Immunologic Deficiency Syndromes Immunosuppression Immunotherapy Interleukin-15 Libraries Malignant Neoplasms Memory Memory impairment Modeling Mouse Strains Mus Nature Nivolumab Organ Pathway interactions Patients Prodrugs Prognosis Proteins RNA Resistance Retroviridae Surface System T cell response T memory cell T-Cell Activation T-Cell Proliferation T-Lymphocyte T-Lymphocyte Subsets Technology Therapeutic Transgenic Mice Viral Virus Work adaptive immune response anti-tumor immune response base cell mediated immune response cohort cytokine exhaustion gene therapy immune checkpoint immune resistance immunogenic cell death immunomodulatory strategy immunoregulation immunosuppressed improved outcome in vivo innovation interleukin-15 receptor ipilimumab knock-down mouse model neoplastic cell new technology novel novel strategies pembrolizumab phase III trial pre-clinical programmed cell death protein 1 resistance mechanism response single cell sequencing systemic toxicity translational potential tumor tumor growth tumor microenvironment

项目摘要

PROJECT SUMMARY/ABSTRACT Antitumor immune responses require a functional repertoire of innate and adaptive immune cells. Glioblastoma (GBM), however, harbors a profoundly immunosuppressed microenvironment, particularly its T cell ignorance caused by bone marrow sequestration; T cell exhaustion caused by immune checkpoint molecules on the surface of T cells that suppress T cell function; and impaired memory T-cell responses. Unfortunately, efforts to target the immunosuppressed GBM microenvironment with systemic immunotherapies have not produced meaningful impact in clinical trials. Localized viral treatments have also been investigated for GBM and, while these viruses elicit an anti-tumoral immune response, these treatments have also failed to impact survival in clinical trials. To address these limitations, we have investigated intratumoral delivery of a replicating retrovirus expressing RLI, which encodes an interleukin-15 fusion protein that enhances CD8+ and CD4+ naïve and memory T-cell proliferation, as a therapeutic strategy free of the toxicities of systemic treatments targeting the tumor microenvironment. We demonstrated that replicating retroviral delivery of RLI prolonged survival of immunocompetent mice with intracranial gliomas using multiple different models. Here, we will build upon our data by investigating our central hypothesis that intratumoral RLI immunomodulatory gene therapy can be potentiated by adding other immunomodulatory strategies, incorporating immunogenic cell death, or targeting resistance mechanisms. We will investigate our hypothesis through four specific aims: (1) Potentiate RLI immunomodulatory gene therapy by enhancing T-cell mobilization, co-stimulation, and memory; (2) Determine if targeting checkpoint pathways potentiates retroviral RLI immunomodulatory gene therapy; (3) Enhance RLI immunomodulatory gene therapy by incorporating immunogenic cell death; and (4) Identify and target glioblastoma-expressed proteins that counteract retroviral RLI immunomodulatory gene therapy. Our pursuit of these aims will utilize novel technologies developed by our lab such as our binary retroviral system to deliver a large payload of immunomodulatory genes and our retroviral compact Cas13d RNA-targeting CRISPR to target resistance mechanisms. We will combine these innovative approaches with cutting-edge technologies such as CyTOF to characterize the effects of RLI-based retroviral therapies on the full cohort of innate and adaptive immune responses; customized CRISPRi libraries; paired immunodeficient and immunocompetent mice strains to isolate immunologic resistance mechanisms; and single cell sequencing to profile T-cell subsets altered by these therapies. These studies will develop our novel localized RLI retroviral immunotherapy in a manner that addresses the spectrum of mechanisms creating local and systemic immunodeficiency in GBM by accounting for T-cell ignorance and exhaustion, and identifying and targeting tumor cell and immune cell-driven resistance mechanisms before they evolve. In doing so, we will validate our hypothesis regarding the impact of RLI-based immunomodulatory gene therapy on GBM, a novel strategy with significant translational potential.

项目摘要/摘要抗肿瘤免疫调查需要先天性和适应性免疫球的功能性曲目。胶质母细胞瘤（GBM）然而，具有深刻的免疫抑制微环境，尤其是其T细胞无知由骨髓隔离引起；由免疫检查点分子引起的T细胞耗尽 T细胞抑制T细胞功能的表面；并损害记忆T细胞响应。不幸的是，努力尚未产生全身免疫疗法的免疫抑制的GBM微环境在临床试验中有意义的影响。还研究了GBM的局部病毒治疗这些病毒引起了抗肿瘤的免疫反应，这些治疗也未能影响临床试验。为了解决这些限制，我们已经研究了重复逆转录病毒的肿瘤内输送表达RLI，该RLI编码介绍CD8+和CD4+幼稚的白介素15融合蛋白记忆T细胞增殖，作为一种治疗策略，没有针对的全身治疗的毒性肿瘤微环境。我们证明了RLI的逆转录病毒延长生存期的复制递送使用多种不同模型的颅内神经胶质瘤的免疫能力小鼠。在这里，我们将建立在我们的基础上通过调查我们的中心假设，即可能是肿瘤内RLI免疫调节基因治疗的数据通过添加其他免疫调节策略，结合免疫原性死亡或靶向来实现电阻机制。我们将通过四个特定目的研究我们的假设：（1）增强RLI 通过增强T细胞动员，共同刺激和记忆，免疫调节基因治疗；（2）确定如果针对检查点途径潜在逆转录病毒RLI免疫调节基因治疗；（3）增强RLI 通过编码免疫原性细胞死亡，免疫调节基因治疗；（4）识别和目标胶质母细胞瘤表达的蛋白质，可抵消逆转录病毒RLI免疫调节基因疗法。我们的追求这些目标将利用我们的实验室开发的新技术，例如我们的二进制逆转录病毒系统来提供免疫调节基因的大量有效载荷和我们的逆转录病毒CAS13D RNA靶向CRISPR 电阻机制。我们将将这些创新方法与诸如尖端技术相结合细胞表征基于RLI的逆转录病毒疗法对整个先天和自适应队列的影响免疫调查；定制的CRISPRI库；配对的免疫缺陷和免疫能力小鼠菌株分离免疫抗性机制；以及对轮廓T细胞测序的T-Cell子集进行了改变这些疗法。这些研究将以一种新型的局部RLI逆转录病毒免疫疗法来发展通过会计解决了在GBM中创造本地和系统的免疫缺陷的机制的范围用于T细胞的无知和疲惫，并识别和靶向肿瘤细胞和免疫细胞驱动的抗性机制在进化之前。在这样做的过程中，我们将验证我们关于基于RLI的影响的假设 GBM上的免疫调节基因治疗，这是一种具有巨大翻译潜力的新型策略。